Orca flash4.0 camera

Images of the metaphase I chromosomes were captured using a DM2000 microscope equipped with a DFC450 camera and controlled by LAS v4.4 system software (Leica Microsystems). For each cell, images were captured in up to 8 different focal planes to aid scoring.
Prophase I meiocytes labeled by FISH and immunofluorescence were optically sectioned using a DM5500B microscope (Leica Microsystems), equipped with a Hamamatsu ORCA-FLASH4.0 camera and controlled by Leica LAS-X software v2.0. Z-stack images of the meiocytes were processed using the deconvolution module of the Leica LAS-X software package. Images were further processed using Fiji (an implementation of ImageJ), a public domain program by W. Rasband available from http://rsb.info.nih.gov/ij/ (Schneider et al., 2012 (link)).

Neurons were fixed and immunolabeled with primary antibodies as described above. After three 5-min washes in PBS, neurons were incubated for 1 h with Alexa Fluor 647-conjugated goat anti-rabbit IgG or IgG subclass-specific anti-mouse secondary antibody (Invitrogen, 1:1500) and CF568-conjugated goat anti-mouse IgG1 secondary antibody (Biotium Cat# 20248, 1:1500). Cells were then subjected to three 5-min washes in PBS, then mounted on glass depression slides in a GLOX-MEA (Tris 10 mM pH 8, glucose oxidase 56 mg/mL, catalase 34 mg/mL, 10 mM MEA) solution. Images were acquired using a Leica Infinity TIRF microscope equipped with a ×163/1.49 NA oil immersion objective and a Hamamatsu ORCA flash 4.0 camera using Leica LASX software for image acquisition. Alexa Fluor‐647 and CF568 images with 40,000–50,000 cycles and an exposure time of 10 ms per channel were collected with the 638 nm and 561 nm excitation lines, respectively.

Hybridization signals in mitotic and meiotic metaphase I spreads were examined using a Leica DM5500B microscope equipped with a Hamamatsu ORCA-FLASH4.0 camera and controlled by Leica LAS X software v2.0. Digital images were processed using Adobe Photoshop CS5 (Adobe Systems Inc., San Jose, CA, USA) extended version 12.0 × 64. Polyacrylamide-embedded meiocytes were optically sectioned using the same Leica DM5500B microscope. Z-stacks were processed using the deconvolution module of the Leica LAS X Software package. Images were processed using Fiji (an implementation of ImageJ, a public domain program by W. Rasband available from http://rsb.info.nih.gov/ij/).

Pollen grains and Pollen Mother Cells stained by the Feulgen technique were imaged using a LEICA DM2000 microscope (Leica Microsystems, http://www.leica-microsystems.com/, accessed on 31 March 2021), equipped with a Leica DFC450 camera and controlled by LAS v4.4 system software (Leica Biosystems, Wetzlar, Germany). Tetrads labelled by FISH were imaged using a Leica DM5500B microscope equipped with a Hamamatsu ORCA-FLASH4.0 camera and controlled by Leica LAS X software v2.0. Z-stacks were processed using the 561-deconvolution module of the Leica LAS X Software package. Images were processed using Adobe Photoshop CS5 (Adobe Systems Incorporated, San Jose, CA, USA) extended version 12.0 × 64.